Recombinant Human HGF (NS0-expressed) GMP Protein, CF

  
  • Purity
    >95%, by SDS-PAGE with silver staining, under reducing conditions.
  • Endotoxin Level
    <0.10 EU per 1 μg of the protein by the LAL method.
  • Activity
    Measured by its ability to induce IL-11 secretion by Saos‑2 human osteosarcoma cells. Hjertner, O. et al. (1999) Blood 94:3883. The ED50 for this effect is 0.1-0.5 ng/mL.
    The specific activity of recombinant human HGF is approximately 1.5 x 103 IU/μg, which is calibrated against recombinant human HGF WHO International Standard (NIBSC code: 96/564).
  • Source
    Mouse myeloma cell line, NS0-derived Gln32-Ser728 Manufactured and tested under cGMP guidelines.
  • Accession #
  • N-terminal Sequence
    Analysis
    Amino acid sequencing of the alpha chain was blocked, suggesting it is consistent with Gln32
    Gln32-Arg-Lys-Arg-Arg-Asn-Thr-Ile-His-Glu ( alpha -chain)
    Val495-Val-Asn-Gly-Ile-Pro-Thr-Arg-Thr-Asn ( beta -chain)
  • Structure / Form
    Disulfide-linked heterodimer
  • Predicted Molecular Mass
    alpha chain: 53.7 kDa beta chain: 26 kDa
  • SDS-PAGE
    65-75 kDa, non-reducing conditions
    60-65 kDa, and 30-40 kDa, reducing conditions
294-GMP
 
Formulation Lyophilized from a 0.2 μm filtered solution in PBS.
Reconstitution Reconstitute at 100 μg/mL in PBS.
Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • A minimum of 6 months when stored at ≤ -20 °C as supplied. Refer to lot specific COA for the Use by Date.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 3 months, ≤ -20 °C under sterile conditions after reconstitution.
Data Images
Recombinant Human HGF GMP induces IL-11 secretion by
Saos‑2 human osteosarcoma cells. The ED50 for this effect is 0.1-0.5 ng/mL.
1 μg/lane of GMP-grade Recombinant Human HGF (Catalog # 294-GMP) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by silver staining, showing R bands at 34-37 and 60-64 kDa and NR bands at 76-83 kDa, respectively.
Background: HGF

HGF, also known as scatter factor and hepatopoietin A, is a pleiotropic protein in the plasminogen subfamily of S1 peptidases. It is a multidomain molecule that includes an N-terminal PAN/APPLE‑like domain, four Kringle domains, and a serine proteinase-like domain that has no detectable protease activity (1‑5). Human HGF is secreted as an inactive 728 amino acid (aa) single chain propeptide. It is cleaved after the fourth Kringle domain by a serine protease to form bioactive
disulfide‑linked HGF with a 60 kDa alpha  and 30 kDa beta  chain. Alternate splicing generates human HGF isoforms that lack the proteinase‑like domain and different numbers of the Kringle domains. Human HGF shares 91%‑94% aa sequence identity with bovine, canine, feline, mouse, and rat HGF. HGF binds heparan-sulfate proteoglycans and the widely expressed receptor tyrosine kinase, HGF R/c-MET (6, 7). HGF-dependent c-MET activation is implicated in the development of many human cancers (8). HGF regulates epithelial morphogenesis by inducing cell scattering and branching tubulogenesis (9, 10). HGF induces the up‑regulation of integrin alpha 2 beta 1 in epithelial cells by a selective increase in alpha 2 gene transcription (11). This integrin serves as a collagen I receptor, and its blockade disrupts epithelial cell branching tubulogenesis (11, 12). HGF can also alter epithelium morphology by the induction of nectin‑1 alpha  ectodomain shedding, an adhesion protein component of adherens junctions (13). In the thyroid, HGF induces the proliferation, motility, and loss of differentiation markers of thyrocytes and inhibits TSH‑stimulated iodine uptake (14). HGF promotes the motility of cardiac stem cells in damaged myocardium (15).

  • References:
    1. Karihaloo, A. et al. (2005) Nephron Exp. Nephrol. 100:e40.
    2. Hammond, D.E. et al. (2004) Curr. Top. Microbiol. Immunol. 286:21.
    3. Rosario, M. and W. Birchmeier (2004) Dev. Cell 7:3.
    4. Lesk, A.M. and W.D. Fordham (1996) J. Mol. Biol. 258:501.
    5. Nakamura, T. et al. (1989) Nature 342:440.
    6. Mizuno, K., et al. (1994) J. Biol. Chem. 269:1131.
    7. Gheradi, E. et al. (2003) Proc. Natl. Acad. Sci. 100:12039.
    8. Corso, S. et al. (2005) Trends Mol. Med. 11:284.
    9. Maeshima, A. et al. (2000) Kid. Int. 58:1511.
    10. Montesano, R. et al. (1991) Cell 67:901.
    11. Chiu, S-J. et al. (2002) J. Biomed. Sci. 9:261.
    12. Saelman, E.U.M. et al. (1995) J. Cell Sci. 108:3531.
    13. Tanaka, Y. et al. (2002) Biochem. Biophys. Res. Commun. 299:472.
    14. Mineo, R. et al. (1994) Endocrinology 145:4355.
    15. Urbanek, K. et al. (2005) Circ. Res. 97:663.
  • Long Name:
    Hepatocyte Growth Factor
  • Entrez Gene IDs:
    3082 (Human); 15234 (Mouse); 403441 (Canine)
  • Alternate Names:
    deafness, autosomal recessive 39; DFNB39; EC 3.4.21; EC 3.4.21.7; fibroblast-derived tumor cytotoxic factor; F-TCF; hepatocyte growth factor (hepapoietin A; scatter factor); Hepatopoeitin-A; Hepatopoietin A; HGF; HGFB; HPTAhepatocyte growth factor; lung fibroblast-derived mitogen; Scatter factor; SF; SFhepatopoeitin-A

GMP Proteins
R&D Systems GMP proteins are produced according to relevant sections of the following documents: WHO TRS, No. 822, 1992 Annex 1, Good Manufacturing Practices for Biological Products; USP Chapter 1043, Ancillary Materials for Cell, Gene and Tissue-Engineered Products and USP Chapter 92, Growth Factors and Cytokines Used in Cell Therapy Manufacturing.

R&D Systems quality focus includes:
  • Manufactured and tested under an ISO 9001:2008 and ISO 13485:2003 certified quality system
  • Documented processes and QA control of documentation and process changes
  • Personnel training programs
  • Raw material testing and vendor qualification/monitoring
  • Fully validated equipment, processes and test methods
  • Equipment calibration schedules using a computerized calibration program
  • Facility maintenance, safety programs and pest control
  • Material review process for variances
  • Monitoring of stability over product shelf-life

R&D Systems strives to provide our customers with the analytical characteristics of each product so that customers may determine whether our products are appropriate for their research. The Certificate of Analysis provided contains the following lot specific information:

  • N-terminal amino acid analysis, SDS-PAGE analysis, and endotoxin level (as determined by LAL assay) performed on each bulk QC lot, not on individual bottlings of each QC lot
  • Post-bottling lot-specific bioassay results (compliance with an established range) and results of microbial bioburden testing (using broth culture, Sabourand'''s dextrose and blood agar plates with results reported at 3 days and at 7 days)

Additional testing and documentation requested by the customer can be arranged at an additional cost. Testing may include, but is not limited to, USP sterility testing, positive identity testing, testing for adventitious agents and testing for residual host cell content.

Production records and facilities are available for examination by appropriate personnel on-site at R&D Systems in Minneapolis, Minnesota USA.

R&D Systems sells its GMP grade recombinant protein products for research use or further manufacturing use in ex vivo cell therapy applications. They are not for in vivo use or for use as therapeutic or other drugs, biologic products or devices. Please read the following End User Terms prior to using this product.

END USER TERMS OF USE OF PRODUCT
The following terms are offered to you upon your acceptance of these End User Terms of Use of Product. By using this product, you indicate your acknowledgment and agreement to these End User Terms of Use of Product. If you do not agree to be bound by and comply with all of the provisions of these End User Terms of Use of Product, you should contact your supplier of the product and make arrangements to return the product.

We suggest you print and retain a copy of these End User Terms of Use of Product for your records.

The End User is aware that R&D Systems, Inc. sells its GMP products for research use only or further manufacturing and not for in vivo use, the production of therapeutics or other drugs or for biologic products or devices. The End User further agrees, as a condition of the sale of R&D Systems GMP products that: a) the End User will not use this GMP Product in any procedure wherein the product may be directly or indirectly administered to humans, unless the End User has obtained, or prior to their use will have obtained, an Investigational New Drug (IND) exemption from the FDA and will use the product only in accordance with the protocols of such IND and of the Institutional Review Board overseeing the proposed research, or b) the End User will use the products outside of the United States in accordance with the protocols of research approved by the Institutional Review Board or authorized ethics committee and regulatory agencies to which the End User is subject to in their territory.

R&D Systems, Inc. has the right, at its sole discretion, to modify, add or remove any terms or conditions of these End User Terms of Use without notice or liability to you. Any changes to these End User Terms of Use are effective immediately following the printing of such changes on this product insert. The most recent version of these End User Terms of Use of Product may be found at: RnDSystems.com/Legal-Information

You agree to review these End User Terms of Use of Product to ensure any subsequent use by you of R&D Systems GMP Products following changes to these End User Terms of Use of Product constitutes your acceptance of all such changes.

TERMS AND CONDITIONS
The following limitation applies to R&D Systems warranty and liability for damages: All products are warranted to meet R&D Systems published specifications when used under normal laboratory conditions.

R&D SYSTEMS DOES NOT MAKE ANY OTHER WARRANTY OR REPRESENTATION WHATSOEVER, WHETHER EXPRESS OR IMPLIED, WITH RESPECT TO ITS PRODUCTS. IN PARTICULAR, R&D SYSTEMS DOES NOT MAKE ANY WARRANTY OF SUITABILITY, NONINFRINGEMENT, MERCHANTABILITY OR FITNESS FOR ANY PARTICULAR PURPOSE.

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Full details of R&D Systems Terms and Conditions of Sale can be found online at: RnDSystems.com/Legal-Information

 
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